Antigen Retrieval Reagent, pH 9 (10X)

Antigen Retrieval Reagent, pH 9 (10X) is used to recover antigens masked by fixation in cross linking fixatives such as formalin.

Product Specification

Formulation:

Liquid. In Tris buffer.

Applications:

IHC

Shipping:

Shipped on Blue Ice

Long Term Storage:

+4°C

Technical Info/Product Notes:

In order to perform immunostaining, tissue specimens should be preserved in an appropriate fixative. Fixation stops tissue autolysis, preserves tissue structures, and immobilizes antigens. However, antigens undergo chemical alteration of their primary, secondary and tertiary structures during fixation. Antigenic sites may be masked due to changes induced in the epitope or neighboring proteins. Enzymatic treatment with proteolytic enzymes (i.e. pepsin, trypsin or pronase) has been performed to expose the masked antigens.

Shi et al. (1991) have reported that treating tissue sections with a heavy metal solution in a microwave oven can cover masked antigens significantly. However, heavy metals in the solution increase the risk of toxic exposure to lab personnel. To reduce the risk, we have developed an antigen unmasking solution which is free of heavy metals. The reagent is provided in a convenient 10X concentrate. Use of this antigen recovery buffer avoids unnecessary heavy metal exposure and other handling and disposal issues.

Interpretation of the results will be the sole responsibility of the user.

Protocol:

Suggested Protocol:

Deparaffinize and rehydrate tissue sections.

Prepare a 1X Antigen Retrieval Solution by mixing 10 mL of Antigen Retrieval Reagent, pH 9 (10X) with 90 mL of deionized water to make 100 mL of solution.

Fill a coplin jar with sufficient Antigen Retrieval Solution (see Step 2) to cover the tissue sections on the slides.

Pre-heat steamer or water bath to 95-100°C.

Place coplin jar in steamer or water bath.

Place deparaffinized slides (1-3 slides/jar) in the coplin jar and incubate for 20-40 min (optimal incubation time and temperature should be determined by the end user, and may vary with different antibodies and tissue types).

Remove coplin jar from the water bath and allow the slides to cool for 20 min to reach room temperature.

Wash slides in deionized water and then with wash buffer. Proceed with immunostaining.